一株抗流产布鲁氏菌疫苗菌株S19的布鲁氏菌噬菌体的分离、电镜及理化性质

D. Chachra, H. Kaur, M. Chandra, H. Saxena
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引用次数: 7

摘要

在印度,布鲁氏菌病是一种经济上重要但被忽视的地方性人畜共患传染病。布鲁氏菌病的患病率从2006-07年的34.15%上升到2010-11年的67.28%。流产布鲁氏菌是一种能够感染动物和人类的细胞内病原体。使用噬菌体治疗细菌感染是现有抗生素治疗的一个有吸引力的替代方案。噬菌体以特定宿主为目标,不太可能在非目标细菌菌株中引起耐药性。从卢迪亚纳市某奶牛场污水样本中分离出一株布氏菌噬菌体,以抵抗处于生长期的流产布氏菌s19菌株。37℃有氧培养48小时后,斑块形态为离散、清晰、直径0.1 ~ 3mm的圆形斑块。野外分离的9株产弧菌对噬菌体敏感。布鲁氏噬菌体的宿主范围是针对疫苗菌株,即B. abortus, B. melitensis和B. suis的S19, S99和Rev1。分离的布鲁氏菌体不能溶解任何外源物种的培养物,包括多杀性巴氏菌、大肠杆菌、葡萄球菌、链球菌、都柏林沙门氏菌、微球菌和假单胞菌。电子显微镜研究表明,该布鲁氏菌体在5万倍放大下约为65 nm,头圆尾短。大小和形状类似于另一种布鲁氏菌噬菌体第比利斯噬菌体和其他地方分离的噬菌体。分离的布鲁氏噬菌体在-20℃、4℃、37℃和50℃的温度下暴露20分钟后能够存活,但在70℃及以上的温度下对布鲁氏噬菌体是致命的。与正常光线不同,阳光对噬菌体生存能力的影响表明了对噬菌体的有害影响。紫外光在15 min内完全破坏噬菌体。非离子型洗涤剂如SDS(10%)在15 min内完全破坏噬菌体。RNAse和胰蛋白酶对噬菌体的生存能力没有影响,而蛋白酶K和溶菌酶降低了噬菌体的生存能力。分离的噬菌体耐pH 7和9,而pH 3和5时噬菌体活性降低。根据所查阅的文献,这是首次报道从旁遮普分离出一种针对流产杆菌的属特异性布鲁氏菌体,这将为其在各种具有成本效益的布鲁氏菌病诊断和治疗中使用铺平道路。
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Isolation, Electron Microscopy And Physicochemical Characterization Of A Brucellaphage Against Brucella Abortus Vaccine Strain S19
Brucellosis is an economically important but neglected emerging endemic zoonotic communicable disease in India. The prevalence of brucellosis has increased from 34.15% of the samples in 2006-07 to 67.28 % in the year 2010-11. Brucella abortus is an intracellular pathogen capable of infecting animals and humans. The use of phages in the treatment of bacterial infections is an attractive alternative to existing antibiotic therapy. Phages target a particular host and it is unlikely to elicit resistance in untargeted bacterial strains. A brucellaphage was isolated against actively growing stage of Brucella abortus strain S 19 from sewage sample of a dairy farm in Ludhiana. The plaque morphology revealed discrete, clear, circular plaques of diameter 0.1 to 3 mm after 48 h of incubation at 37C aerobically. The field isolates (n=9) of B. abortus were sensitive to phage. The host range of brucellaphage is against vaccine strains, viz. S19, S99 and Rev1 of B. abortus, B. melitensis and B. suis. The isolated brucellaphage failed to lyse any culture of heterologous species tested viz. Pasteurella multocida, E.coli, Staphylococcus, Streptococcus, Salmonella Dublin, Micrococcus and Pseudomonas spp. Electron microscopic studies of the brucellaphage revealed it to be an elementary body measuring approximately 65 nm at 50,000 X magnification with rounded head and a very short tail.The size and shape resembles another Brucella phage Tbilisi phage and the other phages isolated elsewhere. The isolated brucellaphage was able to survive at a temperature of -20C, 4C, 37C and 50C when exposed for duration of 20 min. But, a temperature of 70C and beyond was lethal for the brucellaphage. Unlike normal light, the effect of sunlight on the survivability of phage indicates deleterious effects on the phage. UV light completely destroyed the phage within 15 min. Non-ionic detergents like SDS (10%) completely destroyed the phage in 15 min. There was no effect of RNAse and trypsin on the survivability of phage while proteinase K and lysozyme reduced the survivability of the isolated phage. The isolated phage was tolerant to pH 7 and 9 while there was a reduction in phage activity at pH 3 and 5. According to the literature reviewed, this is the first report of isolation of a genus specific brucellaphage against B. abortus from Punjab which will pave a way for its use in various cost effective diagnostics and in therapy of brucellosis.
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